Methods for inducing an immune response against respiratory syncytial virus - Patents.com

Abstract

The present invention provides methods of vaccination and of inducing IgA and cellular immune responses, including T cell immune responses, by administering a deoptimized respiratory syncytial virus (RSV). TIFF2024513917000015.tif98165

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application No. 63 / 172,540, filed April 8, 2021, which is incorporated by reference in its entirety herein.

[0003] Technical Field The present invention relates to vaccines and providing a protective immune response against respiratory syncytial virus (RSV). [Background technology]

[0004] All publications in this specification are incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference herein. The following description contains information that may be useful in understanding the present invention. None of the information provided herein is an admission that it is prior art to or relevant to the invention claimed herein, or that any publication specifically or implicitly mentioned is prior art.

[0005] Respiratory syncytial virus is a non-segmented, single-stranded, negative-stranded, enveloped ribonucleic acid (RNA) virus that belongs to the family Pneumoviridae (formerly Paramyxoviridae). It is transmitted via respiratory secretions by close contact with infected individuals or contact with contaminated surfaces or objects. The humoral immune response usually results in the development of anti-RSV neutralizing antibody titers, but these are often suboptimal during primary infection in infants. The cellular immune response to RSV infection is also important for viral clearance. This immune response is essential for host defense against RSV and is also involved in the immunopathogenesis of severe lower respiratory tract RSV bronchiolitis. The incubation period is 3–5 days, and viral shedding usually occurs for 3–8 days. It is transmitted via respiratory secretions by close contact with infected individuals or contact with contaminated surfaces or objects. Infection can occur when infectious particles come into contact with the mucous membranes of the eyes, mouth, or nose, or when droplets produced by sneezing or coughing are inhaled.

[0006] Although RSV has long been known to be a major pathogen of young children, the threat of RSV in the elderly is becoming recognized. Age-related declines in immune function increase the risk of various infections, including RSV, and community-dwelling elderly people are more susceptible to respiratory illness due to RSV and are at higher risk of subsequent emergency department visits and hospitalizations. A retrospective epidemiological review of 10 clinical trials reporting the incidence rate of laboratory-confirmed RSV in older people attending health care facilities estimated that RSV was responsible for up to 12% of acute respiratory illnesses in older people not selected for comorbidities, with variation across clinical settings and years. The incidence of RSV in health care facilities among older people not selected for having underlying conditions has been observed to increase with age, with 8%–13% of hospitalized adults with chronic cardiopulmonary disease infected with RSV during the winter months. Overall, the duration of hospitalization due to RSV in older people ranged from 3 to 6 days, with a significant proportion of patients requiring intensive care admission and mechanical ventilation. Mortality rates in older people hospitalized with RSV ranged from 6 to 8%.

[0007] Currently, there is no approved RSV vaccine, and there remains a need in the art to provide methods for vaccinating individuals, particularly children and the elderly. Summary of the Invention

[0008] The following embodiments and aspects thereof are described and illustrated in connection with compositions and methods that are intended to be exemplary and illustrative, not limiting in scope.

[0009] Various embodiments of the present invention provide a method of inducing an immune response in a subject, the method comprising intranasally administering to the subject one or more doses of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to induce an immune response, the effective amount being about 10 3 ~10 9 The RSV is deoptimized in focus forming units (FFUs), and the immune response comprises an IgA immune response, a cellular immune response, or both.

[0010] In various embodiments, administering one or more doses of an effective amount of the composition intranasally can include administering two doses, the second dose being administered about 28 days after the first dose. In various embodiments, administering one or more doses of an effective amount of the composition intranasally can include administering two doses, the second dose being administered at least 28 days after the first dose. In various embodiments, administering one or more doses of an effective amount of the composition intranasally can include administering two doses, the second dose being administered about 28-35 days after the first dose.

[0011] In various embodiments, administering intranasally can include administering by nasal drops, hi various embodiments, administering intranasally can include administering by nasal spray.

[0012] In various embodiments, the effective amount is about 10 5 In various embodiments, the effective amount may be about 2×10 5In various embodiments, the effective amount may be about 10 6 In various embodiments, the effective amount may be about 2×10 6 It could be.

[0013] In various embodiments, a single dose of an effective amount can be provided in a volume of about 1 mL. In various embodiments, a single dose of an effective amount can be administered in about 4 aliquots.

[0014] In various embodiments, a first aliquot may be administered to the first nostril, a second aliquot may be administered to the second nostril within 10 minutes after the first aliquot is administered to the first nostril, and a third aliquot and a fourth aliquot may be administered to the first and second nostrils in either order approximately 5-20 minutes after the second aliquot is administered to the second nostril.

[0015] In various embodiments, a first aliquot may be administered to the first nostril, a second aliquot may be administered to the second nostril within 5 minutes after the first aliquot is administered to the first nostril, and a third aliquot and a fourth aliquot may be administered to the first and second nostrils in either order approximately 10-15 minutes after the second aliquot is administered to the second nostril.

[0016] In various embodiments, the cellular immune response can include the development of a T cell response. In various embodiments, the development of a T cell response can be compared to a baseline level of response based on a placebo group or compared to a time point before vaccination of the subject.

[0017] In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from a subject may show at least a 1.3-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from a subject may show at least a 1.4-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from a subject may show at least a 1.5-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from a subject may show at least a 1.6-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from a subject may show at least a 1.7-fold increase in spot forming units (SFU) between baseline values ​​and the highest value at the post-vaccination time point.

[0018] In various embodiments, the effective amount is about 10 6 The FFU and eliciting an immune response can include a statistically significant increase in T cell responses compared to a pre-vaccination time point. In various embodiments, the effective amount is about 2×10 6 Being an FFU and eliciting an immune response can include a statistically significant increase in T cell responses compared to a pre-vaccination time point.

[0019] In various embodiments, the effective amount is about 10 5 The FFU and eliciting an immune response can include an increase in T cell responses compared to a pre-vaccination time point. In various embodiments, the effective amount is about 2×10 5Being an FFU and eliciting an immune response can include an increase in T cell responses compared to a pre-vaccination time point.

[0020] In various embodiments, the effective amount is about 10 6 In various embodiments, the effective amount may be about 2×10 FFU and eliciting an immune response may include at least a 2.4-fold increase in IgA levels in the subject. 6 The antibody may be an FFU, and eliciting an immune response may include at least a 2.4-fold increase in IgA levels in the subject.

[0021] In various embodiments, the effective amount is about 10 5 In various embodiments, the effective amount may be about 2×10 FFU and eliciting an immune response may include at least a 1.1-fold increase in IgA levels in the subject. 5 The antibody may be an FFU, and eliciting an immune response may include at least a 1.1-fold increase in IgA levels in the subject.

[0022] In various embodiments, the effective amount is about 10 5 ~10 6 In various embodiments, the effective amount may be about 2×10 FFU and eliciting an immune response may include at least a 1.3-fold increase in IgA levels in the subject. 5 ~2×10 6 The antibody may be an FFU, and eliciting an immune response may include at least a 1.3-fold increase in IgA levels in the subject.

[0023] In various embodiments, the deoptimized RSV can comprise a genome or antigenome having the sequence of SEQ ID NO:1.

[0024] In various embodiments, the subject may be under 18 years of age. In various embodiments, the subject may be under 13 years of age. In various embodiments, the subject may be under 7 years of age. In various embodiments, the subject may be under 2 years of age. In various embodiments, the subject may be between 18 and 49 years of age. In various embodiments, the subject may be at least 50 years of age.

[0025] Various embodiments provide a method of inducing a T cell immune response in a subject, the method comprising: intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to induce a T cell immune response; and about 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 10 6 or about 2 x 10 6 A deoptimized RSV focus forming unit (FFU), wherein the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting a T cell immune response comprises an increase in the T cell response compared to a pre-vaccination time point.

[0026] Various embodiments provide a method of inducing a T cell immune response in a subject, the method comprising: intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to induce an immune response; and about 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 10 5 or about 2 x 10 5 A deoptimized RSV focus forming unit (FFU), wherein the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting a T cell immune response comprises an increase in the T cell response compared to a pre-vaccination time point.

[0027] Various embodiments provide a method of eliciting an IgA immune response, a cellular immune response, or both in a subject, the method comprising: intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to elicit an immune response; and about 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 2×10 5 A deoptimized RSV for focus forming units (FFU), wherein the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:1, and wherein enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from a subject shows at least a 1.3-fold increase in spot forming units (SFU) between baseline values ​​and the highest value at a time point after vaccination.

[0028] Various embodiments provide a method of eliciting an IgA immune response, a cellular immune response, or both in a subject, the method comprising: intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to elicit an immune response; and about 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 2×10 6 A deoptimized RSV focus forming unit (FFU), wherein the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting an immune response comprises at least a 2.4-fold increase in IgA levels in the subject.

[0029] Other features and advantages of the invention will become apparent from the following detailed description, taken in conjunction with the accompanying drawings, which illustrate, by way of example, various features of embodiments of the invention. [Brief description of the drawings]

[0030] Exemplary embodiments are illustrated in the referenced drawings, in which: It is intended that the embodiments and drawings disclosed herein be considered illustrative and not restrictive.

[0031] [Figure 1A] Total spot counts in ELISPOT for placebo and CodaVax-RSV vaccinated volunteers are shown. The number of SFUs (spot forming units) was normalized to 106 cells and graphed on the Y-axis. [Figure 1B] Total spot counts in ELISPOT for placebo and CodaVax-RSV vaccinated volunteers are shown. The number of SFUs (spot forming units) was normalized to 106 cells and graphed on the Y-axis. [Diagram 2] The change in SFU counts is shown compared to day 1 post-vaccination. The green line indicates a 1.3-fold increase in SFU, which is considered a significant induction of T cell responses. [Diagram 3] T cell responses - geometric mean fold increase in the modified intent-to-treat population. Abbreviations: FFU = focus forming units. GMFR - geometric mean fold increase. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] Description of the invention All references cited herein are incorporated by reference in their entirety as if fully set forth. Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0033] One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. Indeed, the present invention is not limited to the methods and materials described. For purposes of the present invention, the following terms are defined below.

[0034] As used herein, the term "about" when used in conjunction with a referenced numerical indication means the referenced numerical indication plus or minus up to 5% of the referenced numerical indication, unless otherwise specified herein. For example, the language "about 50%" encompasses a range of 45% to 55%. In various embodiments, the term "about" when used in conjunction with a referenced numerical indication can mean the referenced numerical indication plus or minus up to 4%, 3%, 2%, 1%, 0.5%, or 0.25% of the referenced numerical indication, if specifically recited in the claims.

[0035] "Deoptimized" as used herein with respect to viruses such as RSV refers to modified viruses whose genomes or antigenomes have wholly or partially synonymous codons and / or codon rearrangements and / or variations in codon pair bias. This substitution of synonymous codons modifies various parameters, including, for example, reduced codon bias within the genome or antigenome, reduced codon pair bias, density of deoptimized codons and deoptimized codon pairs, RNA secondary structure, CpG dinucleotide content, C+G content, UpA dinucleotide content, translation frameshift sites, translation pause sites, presence or absence of tissue-specific microRNA recognition sequences, or any combination thereof.

[0036] As used herein, "antigenome" refers to the reverse complement of the actual viral genome sequence present inside the virus particle.For example, RSV is a negative-sense virus, and it is common to display genome in antigenomic sense, including the protein coding sequence in the forward direction that is read by ribosomes.

[0037] As used herein, "subject" refers to animal or artificially modified animal.Animals include, but are not limited to, humans, non-human primates, cows, horses, sheep, pigs, dogs, cats, rabbits, ferrets, rodents (e.g., mice, rats, and guinea pigs), bats, snakes, and birds.Artificially modified animals include, but are not limited to, SCID mice with human immune system.In a preferred embodiment, the subject is a human.

[0038] "Prophylactically effective" refers to any amount of a vaccine or virus composition that, when administered to a subject susceptible to a virus infection or a virus-associated disease, induces an immune response that protects the subject from being infected with the virus or from being infected with the disease. "Protecting" a subject means either reducing the subject's chance of being infected with the virus or the subject's chance of developing the disease by at least 2-fold, preferably at least 10-fold, 25-fold, 50-fold, or 100-fold. For example, if a subject has a 1% chance of being infected with a virus, reducing the subject's chance of being infected with the virus by 2-fold will result in a 0.5% chance of the subject being infected with the virus.

[0039] As used herein, "therapeutically effective" is any amount of a vaccine or viral composition that, when administered to a subject suffering from a disease for which the vaccine is effective, induces an immune response in the subject that results in the reduction, remission, or regression of the disorder and / or its symptoms. In preferred embodiments, recurrence of the disorder and / or its symptoms is prevented. In other preferred embodiments, the subject is cured of the disorder and / or its symptoms.

[0040] CodaVax-RSV is based on a live attenuated RSV vaccine strain named MinL4.0, created using the principles of Synthetic Attenuated Virus Engineering (SAVE), a synthetic biology approach to "deoptimize" viral genes to slow translation in human host cells (Coleman 2008; Nouen 2017, incorporated by reference as if fully set forth herein). Attenuation is achieved by extensively recoding viral genes using computer algorithms designed to replace preferred codons and codon pairs with synonymous codons that are translated more slowly. This process is called codon and codon pair deoptimization (CPD). CPD produces hundreds of silent mutations that produce amino acid sequences that are 100% identical to those of the target virus, but attenuate the virus by orders of magnitude. The recoded virus is synthesized de novo. The deoptimized viral genes code for wild-type protein sequences, but CPD causes the host cell ribosomes to produce less of the target protein and at a slower rate. The result is a highly attenuated virus that induces a host cell immune response very similar to that of a wild-type infection. Because the sequence is modified by hundreds to thousands of such deoptimization mutations, the attenuated phenotype of the virus is genetically stable.

[0041] The attenuated RSV strain included in CodaVax-RSV, MinL4.0 (e.g., SEQ ID NO: 1), was synthesized with four additional point mutations in the CPD of the RSV L gene. These four additional sequence changes conferred improved genetic stability, further attenuation, and increased immunogenicity in animal models.

[0042] Thus, provided herein are methods of eliciting an immune response in a subject utilizing MinL4.0 and other similar deoptimized viruses.

[0043] Various embodiments of the present invention provide a method of inducing an immune response in a subject, the method comprising intranasally administering to the subject one or more doses of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to induce an immune response, the effective amount being about 10 3 ~10 9 The RSV is deoptimized in focus forming units (FFUs), and the immune response comprises an IgA immune response, a cellular immune response, or both. In various embodiments, the effective amount is a prophylactically effective amount. In various embodiments, the effective amount is a therapeutically effective amount. In various embodiments, the method induces a protective immune response.

[0044] In various embodiments, the method further includes identifying a subject in need of eliciting an immune response against RSV prior to intranasally administering one or more doses of an effective amount of a composition comprising deoptimized RSV.

[0045] In various embodiments, the immune response is an immune response to RSV. In various embodiments, the immune response is an IgA immune response, a cellular immune response, or both to RSV. In various embodiments, the immune response is a T cell immune response to RSV.

[0046] In various embodiments, administering one or more doses of an effective amount of the composition intranasally comprises administering two doses, the second dose being administered about 28 days after the first dose. In various embodiments, administering one or more doses of an effective amount of the composition intranasally comprises administering two doses, the second dose being administered at least 28 days after the first dose. In various embodiments, administering one or more doses of an effective amount of the composition intranasally comprises administering two doses, the second dose being administered 28-35 days after the first dose. In various embodiments, administering one or more doses of an effective amount of the composition intranasally comprises administering two doses, the second dose being administered 21-49 days after the first dose.

[0047] In various embodiments, the method further comprises administering one or more booster doses one or more years after the initial dosing regimen. For example, 1 year, 2 years, 3 years, 4 years, 5 years, 10 years, 15 years, 20 years, 25 years, 30 years, 35 years, 40 years, 45 years, 50 years, 55 years, 60 years, 65 years, 70 years. In various embodiments, the one or more booster doses are administered when the subject is older, for example, the initial dosing regimen is administered as a child, and the booster dose is administered when the subject is over 50 years old. In various embodiments, the subject is a human subject.

[0048] In various embodiments, administering intranasally includes administering by nasal drops. Nasal drops can be administered via a syringe to aid in proper administration. In various embodiments, administering intranasally includes administering by nasal spray.

[0049] In various embodiments, the effective amount is about 10 4 ~10 8 In various embodiments, the effective amount is about 10 5 ~10 7 FFU deoptimized RSV. For example, about 10 4 ~10 8 FFU or about 10 5 ~10 7 The FFU of deoptimized RSV may be a suitable effective amount in an adult subject. In various embodiments, the effective amount is about 10 3 ~10 6 In various embodiments, the effective amount is about 10 4 ~10 5 FFU deoptimized RSV. For example, about 10 3 ~10 6 FFU or about 10 4 ~10 5 A deoptimized RSV FFU may be an appropriate effective amount in pediatric subjects.

[0050] In various embodiments, the effective amount is about 2-3×10 4 ~108 In various embodiments, the effective amount is about 2-3×10 5 ~10 7 FFU of deoptimized RSV. For example, about 2-3 x 10 4 ~10 8 FFU or approximately 2-3 × 10 5 ~10 7 An FFU of deoptimized RSV may be a suitable effective dose in an adult subject. In various embodiments, the effective dose is about 2-3×10 3 ~10 6 In various embodiments, the effective amount is about 2-3×10 4 ~10 5 FFU of deoptimized RSV. For example, about 2-3 x 10 3 ~10 6 FFU or approximately 2-3 × 10 4 ~10 5 A deoptimized RSV FFU may be an appropriate effective amount in pediatric subjects.

[0051] In various embodiments, the effective amount is that of a single dose.

[0052] In various embodiments, the effective amount is about 10 5 In various embodiments, the effective amount is about 2×10 5 In various embodiments, the effective amount is about 10 6 In various embodiments, the effective amount is about 2×10 6 In various embodiments, the effective amount is the amount of a single dose. For example, the effective amount is about 2×10 6 When FFU and deoptimized RSV is administered as two doses, each dose is approximately 2×10 6 Will have FFU.

[0053] In various embodiments, the effective amount is about 10 3 In various embodiments, the effective amount is about 2×10 3 In various embodiments, the effective amount is about 104 In various embodiments, the effective amount is about 2×10 4 In various embodiments, the effective amount is about 10 5 In various embodiments, the effective amount is about 2×10 5 In various embodiments, the effective amount is about 10 6 In various embodiments, the effective amount is about 2×10 6 In various embodiments, the effective amount is about 10 7 In various embodiments, the effective amount is about 2×10 7 In various embodiments, the effective amount is about 10 8 In various embodiments, the effective amount is about 2×10 8 In various embodiments, the effective amount is about 10 9 In various embodiments, the effective amount is about 2×10 9 It's FFU.

[0054] In various embodiments, a single dose of the effective amount is provided in a volume of about 1 mL. In various embodiments, a single dose of the effective amount is administered in about 4 aliquots. For example, if the effective amount is provided in a volume of 1 mL, the volume of each aliquot will be about 250 μL. In various embodiments, a single dose of the effective amount is provided in about 2 aliquots. In various embodiments, a single dose of the effective amount is provided in about 6 aliquots.

[0055] In various embodiments, a first aliquot is administered to the first nostril, a second aliquot is administered to the second nostril within 10 minutes after the first aliquot is administered to the first nostril, and a third aliquot and a fourth aliquot are administered to the first and second nostrils, in any order, approximately 5-20 minutes after the second aliquot is administered to the second nostril.

[0056] In various embodiments, a first aliquot is administered to the first nostril, a second aliquot is administered to the second nostril within 5 minutes of the first aliquot being administered to the first nostril, and a third aliquot and a fourth aliquot are administered to the first and second nostrils, in either order, approximately 10-15 minutes after the second aliquot is administered to the second nostril.

[0057] In various embodiments, the cellular immune response comprises the development of a T cell response. In various embodiments, the development of the T cell response is compared with a baseline level of response based on a placebo group. In various embodiments, the baseline level of response can be the average level of the placebo group. In various embodiments, the baseline level of response can be the median level of the placebo group.

[0058] In various embodiments, the occurrence of T cell response is compared with the time point before vaccination of the subject.In various embodiments, the method induces an immune response in which T cell response is increased compared with the time point before vaccination.In various embodiments, the increase in T cell response compared with the time point before vaccination is a statistically significant increase.

[0059] In various embodiments, the effective amount is about 10 6 The FFU and eliciting an immune response can include a statistically significant increase in T cell responses compared to a pre-vaccination time point. In various embodiments, the effective amount is about 2×10 6 The FFU and eliciting an immune response may include a statistically significant increase in T cell responses compared to a pre-vaccination time point. In various embodiments, the effective amount is about 10 5 The FFU and eliciting an immune response can include an increase in T cell responses compared to a pre-vaccination time point. In various embodiments, the effective amount is about 2×10 5 Being an FFU and eliciting an immune response can include an increase in T cell responses compared to a pre-vaccination time point.

[0060] The pre-vaccination time point discussed herein can be a time point immediately prior to the first administration of vaccination. For example, T cell responses can be measured from blood drawn on the same day as vaccination, but before vaccination. The pre-vaccination time point discussed herein can be a time point such as about 1 day before vaccination, about 5 days before vaccination, about 7 days before vaccination, about 10 days before vaccination, about 14 days before vaccination, or more. The pre-vaccination time point discussed herein can be a time point such as 1-7 days before vaccination, 1-2 weeks before vaccination, 2-4 weeks before vaccination, 4-8 weeks before vaccination, 2-3 months before vaccination, 3-6 months before vaccination, or more.

[0061] In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.3-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.4-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.5-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.6-fold increase in spot-forming units (SFU) between the baseline value and the highest value at the post-vaccination time point. In various embodiments, enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.7-fold increase in spot forming units (SFU) between the baseline value and the highest value at the post-vaccination time point.

[0062] In various embodiments, the method further comprises performing an enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject to obtain a baseline value. In various embodiments, the method further comprises performing one or more enzyme-linked immunosorbent spot (ELISpot) analyses of peripheral blood mononuclear cells (PBMCs) from the subject post-vaccination.

[0063] In various embodiments, the baseline can be measured from a biological sample obtained before administration of the deoptimized RSV. For example, the biological sample for the baseline measurement is taken on the day of the first administration of the deoptimized RSV, before administration. In another example, the biological sample for the baseline measurement can be taken several days or weeks before the day of the first administration of the deoptimized RSV. As an example, the biological sample for the baseline measurement can be taken as part of identifying a subject that induces an immune response, an IgA immune response, a cellular immune response, or a T cell immune response.

[0064] In various embodiments, the effective amount is about 10 6 In various embodiments, the effective amount is about 2×10 6 Being an FFU and eliciting an immune response includes at least a 2.4-fold increase in IgA levels in the subject.

[0065] In various embodiments, the effective amount is about 10 5 In various embodiments, the effective amount is about 2×10 5 Being an FFU and eliciting an immune response includes at least a 1.1-fold increase in IgA levels in the subject.

[0066] In various embodiments, the effective amount is about 10 5 ~10 6In various embodiments, the effective amount is about 2×10 5 ~10 6 Being an FFU and eliciting an immune response includes at least a 1.3-fold increase in IgA levels in the subject.

[0067] In various embodiments, the increase in IgA levels can be compared to the IgA levels that the subject has before the first dose of deoptimized RSV is administered. In other embodiments, the increase in IgA levels can be compared to the IgA levels that the subject has before the next dose of deoptimized RSV is administered. In various embodiments, these increases in IgA levels are in mucosal IgA.

[0068] In various embodiments, the method further comprises analyzing IgA levels from the subject to obtain a baseline value, hi various embodiments, the method further comprises analyzing IgA levels from the subject one or more times after vaccination.

[0069] In various embodiments, the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO: 1. That is, the deoptimized RSV is encoded by a genome or antigenome having the sequence of SEQ ID NO: 1.

[0070] In various embodiments, the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO: 1, but has up to 12 amino acid substitutions, deletions, or additions. In various embodiments, the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO: 1, but has up to 9 amino acid substitutions, deletions, or additions. In various embodiments, the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO: 1, but has up to 6 amino acid substitutions, deletions, or additions. In various embodiments, the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO: 1, but has up to 3 amino acid substitutions, deletions, or additions. In various embodiments, the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO: 1, but has up to 2 amino acid substitutions, deletions, or additions. In various embodiments, the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO: 1, but has up to 1 amino acid substitution, deletion, or addition.

[0071] In various embodiments, the deoptimized RSV comprises a deoptimized RSV genome or antigenome as described in International Patent Application No. PCT / US2014 / 015274 (published as WO2014 / 124238), which is incorporated by reference herein as if fully set forth. In various embodiments, the deoptimized RSV comprises a deoptimized RSV genome or antigenome as described in International Patent Application No. PCT / US2017 / 053047 (published as WO2018 / 057950), which is incorporated by reference herein as if fully set forth, for example, RSV Min_L-NPM2-1[N88K]L of WO2018 / 057950.

[0072] In various embodiments, the subject is under 18 years old. In various embodiments, the subject is under 17 years old. In various embodiments, the subject is under 13 years old. In various embodiments, the subject is under 7 years old. In various embodiments, the subject is under 6 years old. In various embodiments, the subject is under 2 years old.

[0073] In various embodiments, the subject is between 6 months and 2 years old. In various embodiments, the subject is between 3 and 5 years old. In various embodiments, the subject is between 6 and 11 years old. In various embodiments, the subject is between 12 and 17 years old.

[0074] In various embodiments, the subject is between 18 and 49 years of age. In some embodiments, the subject is at least 50 years of age. In some embodiments, the subject is at least 55 years of age. In some embodiments, the subject is at least 60 years of age.

[0075] Various embodiments provide a method of eliciting an IgA immune response, a cellular immune response, or both in a subject, the method comprising intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to elicit an immune response, and 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 2×10 5and a deoptimized RSV for focus forming units (FFU), the deoptimized RSV comprising a genome or antigenome having the sequence of SEQ ID NO:1, wherein enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.3-fold increase in spot forming units (SFU) between a baseline value and a highest value at a post-vaccination time point. In various embodiments, the effective amount is prophylactically effective. In various embodiments, the effective amount is therapeutically effective. In various embodiments, the method elicits a protective immune response. Various embodiments of the invention provide a method of eliciting an IgA immune response, a cellular immune response, or both in a subject, the method comprising intranasally administering to the subject a first dose of an effective amount of a composition comprising deoptimized respiratory syncytial virus (RSV) to elicit an immune response, the method comprising intranasally administering to the subject a first dose of an effective amount of a composition comprising deoptimized RSV to elicit an immune response, the effective amount being about 2×10 6 The deoptimized RSV of focus forming units (FFUs), the deoptimized RSV comprising a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting an immune response comprises at least a 2.4-fold increase in IgA levels in the subject. In various embodiments, the effective amount is prophylactically effective. In various embodiments, the effective amount is therapeutically effective. In various embodiments, the method elicits a protective immune response.

[0076] In various embodiments, the method further includes identifying a subject in need of eliciting an IgA immune response, a cellular immune response, or both, prior to intranasal administration of one or more doses of an effective amount of a composition comprising deoptimized RSV.

[0077] In various embodiments, the method further comprises performing an enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject to obtain a baseline value. In various embodiments, the method further comprises performing one or more enzyme-linked immunosorbent spot (ELISpot) analyses of peripheral blood mononuclear cells (PBMCs) from the subject post-vaccination.

[0078] In various embodiments, the method further comprises analyzing IgA levels from the subject to obtain a baseline value, hi various embodiments, the method further comprises analyzing IgA levels from the subject one or more times after vaccination.

[0079] In various embodiments, the subject is under 18 years old. In various embodiments, the subject is under 17 years old. In various embodiments, the subject is under 13 years old. In various embodiments, the subject is under 7 years old. In various embodiments, the subject is under 6 years old. In various embodiments, the subject is under 2 years old.

[0080] In various embodiments, the subject is between 6 months and 2 years old. In various embodiments, the subject is between 3 and 5 years old. In various embodiments, the subject is between 6 and 11 years old. In various embodiments, the subject is between 12 and 17 years old.

[0081] In various embodiments, the subject is between 18 and 49 years of age. In some embodiments, the subject is at least 50 years of age. In some embodiments, the subject is at least 55 years of age. In some embodiments, the subject is at least 60 years of age.

[0082] Various embodiments provide a method of inducing a T cell immune response in a subject, the method comprising: intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to induce an immune response; and, 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 10 6and eliciting a T cell immune response comprises an increase in the T cell response compared to a pre-vaccination time point. In various embodiments, the increase is a statistically significant increase. In various embodiments, the effective amount is prophylactically effective. In various embodiments, the effective amount is therapeutically effective. In various embodiments, the method elicits a protective immune response. Various embodiments provide a method of eliciting a T cell immune response in a subject, the method comprising intranasally administering to the subject a first dose of an effective amount of a composition comprising deoptimized respiratory syncytial virus (RSV) to elicit an immune response, and intranasally administering 28-35 days after the first dose, a second dose of an effective amount of a composition comprising deoptimized RSV, the effective amount being about 2×10 6 The deoptimized RSV of focus forming units (FFUs), the deoptimized RSV comprising a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting a T cell immune response comprises an increase in the T cell response compared to a pre-vaccination time point. In various embodiments, the increase is a statistically significant increase. In various embodiments, the effective amount is prophylactically effective. In various embodiments, the effective amount is therapeutically effective. In various embodiments, the method elicits a protective immune response.

[0083] Various embodiments provide a method of inducing a T cell immune response in a subject, the method comprising: intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to induce an immune response; and, 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 10 5The deoptimized RSV of focus forming units (FFUs), the deoptimized RSV comprising a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting a T cell immune response comprises an increase in the T cell response compared to a time point prior to vaccination. In various embodiments, the effective amount is prophylactically effective. In various embodiments, the effective amount is therapeutically effective. In various embodiments, the method elicits a protective immune response.

[0084] Various embodiments provide a method of eliciting a T cell immune response in a subject, the method comprising: intranasally administering to the subject a first dose of an effective amount of a composition comprising a deoptimized respiratory syncytial virus (RSV) to elicit an immune response; and, 28-35 days after the first administration, intranasally administering a second dose of an effective amount of the composition comprising the deoptimized RSV, the effective amount being about 2×10 5 The deoptimized RSV of focus forming units (FFUs), the deoptimized RSV comprising a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting a T cell immune response comprises an increase in the T cell response compared to a time point prior to vaccination. In various embodiments, the effective amount is a prophylactically effective amount. In various embodiments, the effective amount is a therapeutically effective amount. In various embodiments, the method elicits a protective immune response.

[0085] In various embodiments, the method further includes identifying a subject in need of eliciting a T cell immune response prior to intranasal administration of one or more doses of an effective amount of a composition comprising deoptimized RSV.

[0086] In various embodiments, the method further comprises performing an enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject to obtain a baseline value. In various embodiments, the method further comprises performing one or more enzyme-linked immunosorbent spot (ELISpot) analyses of peripheral blood mononuclear cells (PBMCs) from the subject post-vaccination.

[0087] In various embodiments, the method further comprises analyzing a T cell immune response from the subject to obtain a baseline value, hi various embodiments, the method further comprises analyzing a T cell immune response from the subject one or more times after vaccination.

[0088] In various embodiments, the subject is under 18 years old. In various embodiments, the subject is under 17 years old. In various embodiments, the subject is under 13 years old. In various embodiments, the subject is under 7 years old. In various embodiments, the subject is under 6 years old. In various embodiments, the subject is under 2 years old.

[0089] In various embodiments, the subject is between 6 months and 2 years old. In various embodiments, the subject is between 3 and 5 years old. In various embodiments, the subject is between 6 and 11 years old. In various embodiments, the subject is between 12 and 17 years old.

[0090] In various embodiments, the subject is between 18 and 49 years of age. In some embodiments, the subject is at least 50 years of age. In some embodiments, the subject is at least 55 years of age. In some embodiments, the subject is at least 60 years of age.

[0091] The pre-vaccination time point discussed herein can be a time point immediately prior to the first administration of vaccination. For example, T cell responses can be measured from blood drawn on the same day as vaccination, but before vaccination. The pre-vaccination time point discussed herein can be a time point such as about 1 day before vaccination, about 5 days before vaccination, about 7 days before vaccination, about 10 days before vaccination, about 14 days before vaccination, or more. The pre-vaccination time point discussed herein can be a time point such as 1-7 days before vaccination, 1-2 weeks before vaccination, 2-4 weeks before vaccination, 4-8 weeks before vaccination, 2-3 months before vaccination, 3-6 months before vaccination, or more. EXAMPLES

[0092] The following examples are provided to better illustrate the claimed invention and should not be construed as limiting the scope of the invention. To the extent that specific materials are mentioned, they are merely for illustration and are not intended to limit the invention. Those skilled in the art may develop equivalent means or reactants without the exercise of inventive capacity and without departing from the scope of the invention.

[0093] Example 1 The following clinical trial protocol was used to generate the data described in Examples 2 and 3. This protocol includes inclusion criteria, exclusion criteria, medications, contraception, and other restrictions, but these criteria and restrictions should not be construed as limitations on the practice of the claimed invention unless specifically excluded in the claims. Indeed, certain individuals excluded from this clinical trial are contemplated as individuals who can and should receive the methods described herein.

[0094] CodaVax-RSV is a sterile solution administered via IN administration. In this FIH Phase I clinical trial in healthy older adults, approximately 2 × 10 5 and 2 x 10 6 Two nominal dose levels of CodaVax-RSV in FFU will be administered 28 days apart (i.e., on days 1 and 29). Subjects will be randomly assigned 2:1 within each group to CodaVax-RSV or saline placebo. The dose levels studied are shown in Table 1.

[0095] The decision to continue dosing after the Sentinel dose and to escalate dose to higher dose cohorts will depend on review of new safety data by the SMC. The PI will review safety data from the first day of dosing for the first two Sentinels and advise the remaining SMC before dosing the remaining Sentinel cohorts. The SMC will then review blinded safety data through Day 8 for all Sentinel and low-dose cohort participants, and then review available blinded safety data before randomizing subsequent participants in each group and before each cohort participant begins receiving their second dose.

[0096] [Table 1]

[0097] The clinical trial will be conducted on healthy male and female volunteers aged 18-49 years (at the time of informed consent) in the sentinel group and 50-75 years (at the time of informed consent) in the main treatment group.

[0098] Women of childbearing potential will be included and will be required to use contraception for the duration of the study, from screening through 6 months after the second vaccination. Women of childbearing potential must have a negative pregnancy test at screening and before receiving the study drug, in accordance with the regulatory Guidance on Nonclinical Safety Studies for the Conduct of Human Clinical Trials and Marketing Authorization for Pharmaceuticals (U.S. Food and Drug Administration [FDA] 2006). TIFF2024513917000003.tif242165TIFF2024513917000004.tif99165

[0099] Overall study design The study is a phase 1, randomized, double-blind, placebo-controlled trial evaluating the safety, tolerability, and immunogenicity of two doses of CodaVax-RSV monovaccination administered IN, 28 days apart. The trial will include two sentinel groups of six young adults aged 18-49 years, followed by approximately 36 healthy older adults aged 50-75 years with low baseline RSV titers.

[0100] Eligible participants will be randomly assigned 2:1 to receive CodaVax-RSV or placebo (saline) in two dose-escalation cohorts. All participants will receive two doses, 28 days apart. The low-dose cohort will receive approximately 2 × 10 5 Nominal dose of FFU, approximately 2 × 10 for the high-dose cohort 6 A nominal dose of FFU will be administered IN, 250 µL in each nostril, repeated 10-15 min later, for a total volume of 1 mL. Dose levels and administration for this trial are as follows (Table 2):

[0101] [Table 2]

[0102] After providing informed consent, potentially eligible participants will be screened by routine physical examination, ECG, and laboratory tests (hematology, biochemistry, and urinalysis) performed within 28 days prior to randomization to determine eligibility. Elderly participants will be recruited from a previous screening study designed to identify individuals with low baseline RSV antibody titers. On Day 1, after physical examination, blood draw for PBMC and serum antibodies, and NP swab for mucosal antibodies, eligible participants will be randomly assigned to either active treatment or placebo within each dose group using a block randomization design provided to the on-site pharmacist by an unblinded statistician for dose groups defined as in Table 6 above. Participants will receive the first of two vaccines, be observed for 2 hours post-dose, and be discharged after successful completion of all specified assessments.

[0103] Participants will complete a daily diary recording oral temperature, presence and severity of involuntary symptoms for 7 days after each vaccination. Participants will return for targeted physical examinations, serum collection, NP swabs and AEs, and concomitant medication reviews according to the assessment schedule. On day 8, safety laboratory tests and PBMC collection will also be performed. Participants will receive a second vaccination.

[0104] Twenty-eight days after the first vaccination (Day 29). Participants will be observed for two hours after dosing and will be discharged after successful completion of all designated evaluations. Study procedures on Days 32, 36, and 43 will be similar to those following the first vaccination. On Day 57, participants will undergo similar evaluations with the addition of a follow-up pregnancy test and ECG.

[0105] Oversight of the trial will be provided by the SMC, which will be comprised of the investigator, the sponsor's MM, and an independent MM. The PI will review safety data from the first day of dosing for the first two Sentinels and advise the remaining SMC before dosing the remaining Sentinel cohorts. The SMC will then review blinded safety data through Day 8 for all Sentinel participants and the low-dose group before randomizing participants to subsequent groups, and will review available blinded safety data before each cohort participant begins receiving their second dose. Two late follow-up visits are scheduled after Day 57.

[0106] Participants who develop symptoms of respiratory infection during the late follow-up period will have a specimen collected for a respiratory viral PCR panel (followed by sequence confirmation if RSV positive), which may be during an unscheduled visit. Clinically significant laboratory abnormalities may also be repeated at an unscheduled visit. All AEs and concomitant medications will be recorded through Day 57, and MAEs, NCIs, SAEs, and concomitant medications of special note (vaccines and immunosuppressants) will be recorded from Day 1 through Day 209 (EOS). Study visits and evaluations will occur as described in the evaluation schedule.

[0107] Number of participants Approximately 42 participants will be randomized.

[0108] Treatment Allocation Randomization Clinical Network Services will generate the randomization schedule request and manage the randomization of eligible study participants according to this document. The randomization list will be generated using a statistical software package by NVT Biostatistician.

[0109] Each participant will be provided with a unique screening number after documentation of informed consent. Once determined eligible to participate in the study, participants will be assigned a sequential randomization number prior to the first dose. Participants who withdraw from the study without completing all required screening assessments, for any reason, will be considered as screening failures.

[0110] Details of the randomization will be further documented in the request for a randomization schedule.

[0111] Safety monitoring committee Oversight will be provided by an SMC, including at least the PI, the sponsor's MM, and an independent MM, which will conduct a review of blinded safety data from the sentinel cohort prior to randomization of the remaining participants in the low-dose cohort, prior to dose escalation to the high-dose cohort, and before participants receive the second vaccination in each group.

[0112] After each review, the SMC will make the following recommendations: Continue clinical trials as planned -Continue the trial with changes Temporarily suspend or terminate a clinical trial If a decision is made to temporarily suspend or terminate a clinical trial, the relevant SMC minutes will be submitted to the HREC.

[0113] Stopping Criteria If any of the following stopping rules are met, randomization and dosing will be halted until the SMC conducts a blinded safety data review to assess relevance and risk to subsequent study participants. Potentially relevant SAEs Grade 4 laboratory values ​​or AEs Two or more similar grade 3 events or laboratory abnormalities. Records of such blinded reviews will be submitted to the HREC.

[0114] Criteria for Completing a Clinical Trial The trial will be completed as planned unless: New information or other evaluation of the safety of the investigational medicinal product indicates a change in the known risk / benefit profile of the compound such that the risk / benefit is no longer acceptable for participants entering the trial. This may be determined by the sponsor, the investigator, the local MM, SMC, HREC, or the regulatory authority. The trial is terminated by the sponsor for administrative reasons. If the sponsor, HREC or regulatory authority chooses to terminate or discontinue the conduct of the trial or the participation of the sites in the trial, the sponsor will provide protocol-specific procedures for early termination or discontinuation, which will be carried out by the sites during the termination or trial halt.

[0115] Participant selection criteria To be eligible for this trial, participants must meet all of the following inclusion criteria: 1. Healthy male and female volunteers aged 50-75 years (at the time of informed consent) in the main treatment group, or 18-49 years (at the time of informed consent) in the sentinel group. 2.BMI ≥ 18.0kg / m2 and ≤ 35kg / m2. 3. Participants must be willing to comply with the following conditions in order to prevent the spread of GMOs in accordance with the OGTR license (DIR161): a. Hygiene measures aimed at preventing person-to-person transmission of the investigational product must be implemented, including, but not limited to, frequent handwashing with soap or hand sanitizer, respiratory hygiene, and cough etiquette within 14 days after each vaccination. b. Do not donate blood, tissue, or organs from the time of your first vaccination until 6 months after your last vaccination. c. Avoid contact with exempt individuals*, children under 2 years of age, and residents of elderly care facilities for 14 days after receiving each vaccine. d. All tissues and materials used to collect respiratory secretions for 14 days after each vaccination must be sealed in a primary container, placed in a secondary container, and made inaccessible to children and animals until returned to the investigational site for disposal. *Exclusions include women who are pregnant or may be pregnant, and individuals with immunodeficiencies or immunosuppression resulting in complete or partial impairment of the immune system. 4. Adequate venous access in case of repeated phlebotomy. 5. Screening test results are within the normal range, or grade 1 abnormality if documented by the investigator to be clinically non-significant. Creatine kinase or bilirubin may be grade 2 if associated with normal ALT and AST and the investigator deems the results clinically non-significant due to strenuous exercise or Gilbert's syndrome. 6. Screen negative for drugs and alcohol (unless explained by prescribed medications). 7. For participants in the primary treatment arm, qualifying RSV neutralizing antibody titers as defined by a previous sero-epidemiology study. 8. Females of childbearing potential must be non-pregnant and non-lactating and must use an acceptable, highly effective dual barrier method of contraception from screening until 6 months after final vaccination. A dual barrier method is defined as a condom and one other method of contraception as follows: · Established hormonal contraception methods (OCPs, long-acting implantable hormones, injectable hormones). · Vaginal ring or IUD. Documented evidence of surgical sterilization at least 6 months prior to screening (e.g., tubal occlusion, hysterectomy, bilateral salpingectomy, or bilateral oophorectomy for females, or vasectomy for male partners if they are the only partners [with appropriate documentation of post-vasectomy showing absence of sperm in semen]). Women who are not of childbearing potential must be at least 12 months postmenopausal. Postmenopausal status is determined by FSH levels of 40 IU / mL or greater at screening for amenorrhea-prone female participants. Women who are abstaining from heterosexual intercourse are also eligible. Periodic abstinence methods (eg, calendar, ovulation, symptothermal, postovulation methods) and pull-out methods are not considered to be highly effective methods of birth control. Female participants in same-sex relationships are not required to use contraception. WOCBP must have a negative pregnancy test at Screening and Day 1 and be willing to undergo additional pregnancy tests as needed throughout the study. Men must be surgically infertile (>30 days post-vasectomy and no viable sperm) or abstinent, or if in a sexual relationship with a WOCBP, their partner must be surgically infertile (e.g., tubal occlusion, hysterectomy, bilateral salpingectomy, bilateral oophorectomy) or use an acceptable highly effective method of double barrier contraception from screening until 6 months after final vaccination. Acceptable methods of double barrier contraception include the use of condoms and the use of effective contraceptives for the female partner, including OCPs, long-acting implantable hormonal, injectable hormonal, vaginal ring or IUD. Participants with same-sex partners (who abstain from penile-vaginal intercourse) are eligible to participate if this is their preferred and usual lifestyle practice. 9. Men should not donate sperm for at least 6 months after their final vaccination. 10. Participants must be able and willing to attend required visits to the CRU. 11. Participant must be willing and able to provide written informed consent prior to the start of any study procedures.

[0116] Participant Exclusion Criteria Participants who meet any of the following exclusion criteria must be excluded from the trial. 1. Pregnant or breastfeeding at the time of screening, or planning pregnancy (by yourself or your partner) within 6 months of receiving your last vaccine. 2. Household contacts or caregivers of children under 2 years of age or immunocompromised individuals (for up to 14 days after last vaccination). Immunocompromised individuals are defined as, but not limited to: a. HIV infected person b. Those who have undergone chemotherapy within the past 6 months c. Those receiving immunosuppressants d. People living with solid organ or bone marrow transplants 3. Positive screening result for HIV, HBV, or HCV 4. Asthma or other chronic lung disease of more than mild severity. Participants will be specifically excluded if they have a history of any of the following related to asthma or lung disease: A. Symptoms have occurred daily for more than one week in the past five years; b. Use of short-acting beta-2 agonists (e.g., albuterol) in the past 5 years; c. Use of inhaled steroids, theophylline or pulsed systemic steroids in the past 5 years, or d. History of intubation or hospitalization related to asthma or other chronic pulmonary disease. 5. History of diabetes (gestational diabetes is permitted if no treatment is required after delivery and serum glucose levels are currently within normal range) 6. History of coronary artery disease, arrhythmia, or congestive heart failure. 7. Clinically significant ECG abnormalities as determined by the investigator at screening. 8. Uncontrolled hypertension (systolic blood pressure >150mmHg or diastolic blood pressure >95mmHg) at screening or prior to dosing on Day 1. 9. History of anaphylaxis or angioedema. 10. History of severe reactions to vaccinations. 11. Known allergy to any component of the vaccine formulation. 12. History of chronic rhinitis, defective nasal septum, cleft palate, nasal polyps, or other nasal abnormalities that may alter the nasal mucosa and affect vaccine response. 13. Have had nasal surgery or nasal cauterization in the past. 14. Have epistaxis (nosebleed), symptoms of upper respiratory tract infection, or subjective fever or fatigue within 3 days prior to dosing on Day 1 (temporary exclusion criteria). 15. Any symptoms or signs on Day 1 that may interfere with proper administration of IP or interpretation of the diary for involuntary AEs (e.g., temperature >38°C, nasal congestion or rhinorrhea) (temporary exclusion criteria) 16 Known or suspected malignancy, except for non-melanoma skin cancer and other early stage surgically resected malignancies that the investigator believes to have a very low chance of recurrence. 17. Participant with immune deficiency, including use of corticosteroids (including IN steroids), alkylating agents, antimetabolites, radiation, immunomodulatory biologics, or other immunomodulatory therapies within 90 days prior to dosing on Day 1, or who plans to use any of these during the follow-up period. Use of topical steroid creams on the skin is permitted. 18. History of bleeding disorders (e.g., factor deficiencies, coagulation disorders, or platelet disorders requiring special precautions). 19. History of autoimmune or demyelinating disease. 20. History of psychiatric illness, psychiatric hospitalization, or suicide attempt within the past 10 years. 21. History of seizures (excluding childhood febrile convulsions), dementia or progressive neurological disease. 22. Received IN medications (including OTC medications but excluding saline) within 30 days prior to Day 1. 23. Have received any other IP within 30 days or 5 half-lives (whichever is longer) prior to Day 1. 24. Have had a vaccination, TB skin test, or allergy vaccination within 30 days prior to Day 1, or are scheduled to receive one by Day 57. 25. Received IN vaccination within 90 days prior to Day 1. 26. Receiving a licensed or investigational RSV vaccine. 27. Having received a blood transfusion or blood products within 90 days prior to Day 1, or scheduled to receive one by Day 57. 28. Planned hospitalization or surgery is scheduled by day 57. 29. Any other chronic medical conditions not listed must be stable without requiring a medication change within the prior 30 days from Day 1. 30.IN Past or current use of illicit drugs. 31. Smokers of any kind (cigarettes, e-cigarettes, marijuana, etc.). History smokers must have quit smoking at least 30 days prior to Day 1. 32. Residents of elderly care facilities 33. Employees of Codagenix, vendors, or clinical trial sites associated with a clinical trial. 34. Any medical, psychiatric or social condition, or professional or other responsibilities, that, in the investigator's judgment, would interfere with or contraindicate the participant's ability to comply with the clinical trial protocol, evaluate safety (including reactogenicity), or give informed consent. 35. A positive PCR test result for COVID-19 within 28 days, or contact with a person with known or suspected COVID-19, or travel to an area with community transmission of COVID-19 within 14 days prior to randomization.

[0117] Medication, birth control, and other restrictions dosage The following drugs are prohibited until day 57: Systemic, intranasal or inhaled steroids Systemic, intranasal, or inhaled immunosuppressants Intranasal vaccines or medications (other than saline) Any kind of vaccine, or skin tests for TB or allergies Blood transfusion or immunoglobulin treatment Other IPs

[0118] NOTE: Antipyretics and analgesics may be used to manage fever or other symptoms after daily recording of involuntary events and temperature and will be recorded as concomitant medications, but should not be administered prophylactically.

[0119] The use of investigational vaccines or other biologics against RSV is prohibited throughout the trial.

[0120] Immunosuppressants Immunosuppressant medications (including but not limited to the list below) are prohibited from 90 days prior to Day 1 through the end of the study. The following immunosuppressant concomitant medications are considered to be of special interest for the purposes of this study: Corticosteroids Prednisone (Deltasone, Orasone) Budesonide (Entocort EC) Prednisolone (Millipred) Calcineurin Inhibitors Cyclosporine (Neoral, Sandimmune, SangCya) Tacrolimus (Astagraf XL, Envarsus XR, Prograf) Target of action of rapamycin (mTOR) inhibitors Sirolimus (Rapamune) Everolimus (Afinitor, Zortress) Inosine monophosphate dehydrogenase (IMDH) inhibitors Azathioprine (Azasan, Imuran) Leflunomide (Arava) Mycophenolic acid (CellCept, Myfortic) Biologics Abatacept (Orencia) Adalimumab (Humira) Anakinra (Kineret) Certolizumab (Cimzia) Etanercept (Enbrel) Golimumab (Simponi) Infliximab (Remicade) Ixekizumab (Taltz) Natalizumab (Tysabri) Rituximab (Rituxan) ·Secukinumab (Cosentyx) Tocilizumab (Actemra) Ustekinumab (Stelara) Vedolizumab (Entyvio) Monoclonal antibodies Basiliximab (Simulect) Daclizumab (Zinbryta) Muromonab (Orthoclone OKT3)

[0121] Additional concomitant medications of note The following agents would be considered concomitant agents of particular interest for the purposes of this protocol: ·vaccine Blood products Immunoglobulin products

[0122] smoking Smokers of any kind (cigarettes, e-cigarettes, marijuana, etc.) are excluded from this study. Previous smokers must have quit smoking at least 30 days prior to Day 1.

[0123] contraception Contraceptive requirements are as described in this example.

[0124] Explanation of the investigational drug CodaVax-RSV CodaVax-RSV is composed of a single-stranded negative-sense RNA [(-)ssRNA] generated by a CPD open reading frame (ORF) designed using computational algorithms (Coleman 2008; Mueller 2010) and is based on the RSV sequence M74568, the biological wild-type RSV strain A2 (Nouen 2017).

[0125] CodaVax-RSV is delivered IN via a syringe (without a needle). The vaccine in a vial is thawed and loaded into a syringe by the clinic's pharmacy (in a class II biosafety cabinet) prior to IN administration. A lower test dose is achieved by on-site dilution using Leibovitz's L-15 medium. Further details are provided in the pharmacy manual.

[0126] placebo The placebo in this trial is saline delivered IN via syringe (no needle).

[0127] Concomitant medications All medications taken in the 30 days prior to screening, including over-the-counter medications, vitamins, and herbal supplements, will be recorded and reviewed by the investigator to determine whether the participant is suitable to take part in the study.

[0128] Use of IP or investigational devices within 30 days prior to screening is prohibited.

[0129] Prior treatment or concomitant therapy (after administration of the investigational product) with any medications, both prescription and non-prescription, must be discussed with the investigator and sponsor's MM prior to administration of the investigational product, unless necessary to treat an AE or appropriate medical treatment needs to be initiated before the investigator consults with the independent MM.

[0130] Paracetamol / acetaminophen (1–2 therapeutic doses per week) may be used for minor illness during the study, at the investigator's discretion and without prior consultation with the sponsor's MM.

[0131] All medications taken by participants through Day 57, including over-the-counter drugs, vitamins, and herbal supplements, will be recorded on the eCRF and coded using the most current WHO Drug Dictionary available in the NVT. After Day 57, only concomitant medications of special note will be recorded. Previous medications and concomitant medications will be listed for each participant and organized by Anatomical Therapeutic Chemical (ATC) and PT.

[0132] Blinding The blinding procedures for the trial will be described in detail in the Data Management Plan and the Pharmacy Manual.

[0133] The sponsor, investigator, MM, investigators, and participants should not attempt to identify which treatment is being received. This study will use unblinded pharmacy (or other qualified facility) personnel to prepare the investigational product.

[0134] In emergency situations, only when knowledge of the investigational product is essential for the clinical management or welfare of a particular participant, the investigator may unblind a participant's treatment assignment. A copy of the randomization sequence and treatment code will be kept in the study site pharmacy. If necessary, the investigator may obtain a participant's treatment assignment from an unblinded member of the pharmacy staff. However, before unblinding, the investigator is strongly encouraged to discuss options with the MM, sponsor, or appropriate study personnel. The investigator will record in source documents the date and reason for revealing the participant's blinded treatment assignment, and the name and role of the person who was unblinded.

[0135] The investigator must notify the sponsor as soon as possible if blinding is broken for any reason and the investigator is unable to contact the sponsor before blinding is broken, without revealing the participant's investigational treatment assignment (unless it is important for the safety of the participant remaining in the trial).

[0136] If the investigator determines that the AE is serious enough to require immediate and specific knowledge of the identity and dose of the product involved, the investigator may release the participant's study code.

[0137] All blinding events must be promptly reported to the MM, the sponsor, and the CNS.

[0138] A soft lock will be placed on the database after the day 57 visit, at which point the statistical team will unblind and generate interim tables and figures. The study sites, study monitors, medical monitors, and sponsors will remain blinded to individual treatment allocations until the end of the study.

[0139] Storage of investigational drugs Once received, the investigational product must be stored at -80°C ± 10°C.

[0140] Filled syringes should be stored at 5 ± 3 °C for no more than 4 hours before use and transported on ice to the clinic.

[0141] The Investigator or designee is solely responsible for the security, accessibility and storage of the investigational product at the investigational site.

[0142] Preparation of investigational drug Procedures for preparation and dispensing of study medication are outlined in the pharmacy manual.

[0143] Administration The investigator or designee is responsible for training study staff and participants on the correct administration of the investigational drug. Doses will be administered in a volume of 1 mL, 250 μL in each nostril, repeated 10-15 minutes after administration to the second nostril. At each application, the second nostril should be administered within 5 minutes of the first nostril.

[0144] Clinical trial schedule When possible, evaluations scheduled at the same evaluation time point should be performed in order from least invasive to most invasive.

[0145] After vaccination Participants will remain in the CRU until all scheduled post-dose procedures are completed. The following steps will occur after administration: · Focused physical examination 2 hours (± 30 minutes) after administration. Vital signs (systolic and diastolic blood pressure, respiratory rate, pulse rate, oral temperature) 2 hours (± 30 minutes) after dosing (after participants had rested in a supine position for at least 5 minutes)

[0146] NOTE: AEs and concomitant medications will be monitored continuously until the participant is discharged from the CRU.

[0147] Participants will be provided with a diary card and a thermometer and instructions on how to record oral temperatures and reactogenic events.

[0148] Participants will be discharged approximately two hours after administration, after all evaluations and procedures have been successfully completed.

[0149] Telephone follow-up (day 2) Participants will be contacted by site staff, who will maintain a diary and ask about any AEs or concomitant medication use. Participants who report grade 3 symptoms of respiratory viral infection will be asked to attend an unscheduled visit and provide a NP swab for PCR.

[0150] Follow-up (Day 4 and Day 8 [± 1 day]) Participants will return to the CRU for follow-up visits on days 4 and 8 (± 1 day).

[0151] Unless otherwise instructed, the following steps will be performed at each follow-up visit: ·Intensive physical examination ·NP swabs for vaccine shedding assessment Vital signs (systolic and diastolic blood pressure, respiratory rate, pulse rate, oral temperature) (after participants had rested in a supine position for at least 5 minutes) Blood sampling for clinical tests (hematology, biochemistry) (only on the 8th day) -Blood sampling to assess viremia in serum Blood collection to assess T cell responses in PBMCs (day 8 only) Review of diaries for reactogenic events Review of other AEs and concomitant medications Diary cards will be collected at the fourth visit (day 8 [± 1 day]).

[0152] Follow-up (15th day [± 3 days]) Participants will return to the CRU for a follow-up visit on Day 15 (± 3 days). The following steps will be performed at the follow-up visit: ·Intensive physical examination ·NP swabs for evaluation of vaccine shedding and humoral immunogenicity (IgA antibodies) Blood sampling to assess immunogenicity (antibodies in serum) Review of other AEs and concomitant medications

[0153] Date of second vaccination (day 29 [± 3 days]) Before vaccination Participants will present to the CRU and, unless otherwise specified, the following assessments will be performed within 2 hours prior to study drug administration: · Screening to determine whether the participant remains eligible to participate in the study Review of other AEs and concomitant medications Urine drug testing and breath alcohol testing · Urine pregnancy test (WOCBP only) Urine testing ·NP swabs for evaluation of vaccine shedding and humoral immunogenicity (IgA antibodies) Vital signs (systolic and diastolic blood pressure, respiratory rate, pulse rate, oral temperature) (after participants had rested in a supine position for at least 5 minutes) Blood sampling for clinical tests (hematology, biochemistry) Blood sampling to assess immunogenicity (antibodies in serum) Blood collection to assess T cell responses in PBMCs

[0154] Vaccination Participants will receive the study drug according to their assigned treatment. Study drug will be administered by assigned clinical staff.

[0155] After vaccination Participants will remain in the CRU until all scheduled post-dose procedures are completed. The following steps will occur after administration: -Intensive physical examination 2 hours (± 30 minutes) after administration Vital signs (systolic and diastolic blood pressure, respiratory rate, pulse rate, oral temperature) 2 hours (± 30 minutes) after dosing (after participants had rested in a supine position for at least 5 minutes)

[0156] NOTE: AEs and concomitant medications will be monitored continuously until the participant is discharged from the CRU.

[0157] Participants will be issued with a new diary card and given instructions on how to record oral temperatures and reactogenic events.

[0158] Participants will be discharged approximately two hours after administration, after all evaluations and procedures have been successfully completed.

[0159] Telephone follow-up (day 30) Participants will be contacted by site staff, who will inquire about any AEs or concomitant medication use. Participants who report grade 3 symptoms of respiratory viral infection will be asked to attend an unscheduled visit and provide a NP swab for PCR.

[0160] Follow-up (Days 32 and 36 [± 1 day]) Participants will return to the CRU for follow-up visits on days 32 and 36 (± 1 day).

[0161] Unless otherwise instructed, the following steps will be performed at each follow-up visit: ·Intensive physical examination ·NP swabs for vaccine shedding assessment Vital signs (systolic and diastolic blood pressure, respiratory rate, pulse rate, oral temperature) (after participants had rested in a supine position for at least 5 minutes) Blood sampling for clinical tests (hematology, biochemistry) (only on the 36th day) -Blood sampling to assess viremia in serum Blood collection to assess T cell responses in PBMCs (day 36 only) Review of diaries for reactogenic events Review of other AEs and concomitant medications. Diary cards will be collected at Visit 8 (Day 36 [± Day 1]).

[0162] Follow-up (Day 43 [± 2 days]) Participants will return to the CRU for a follow-up visit on Day 43 (± 2 days). The following steps will be performed at the follow-up visit: ·Intensive physical examination ·NP swabs for evaluation of vaccine shedding and humoral immunogenicity (IgA antibodies) Blood sampling to assess immunogenicity (antibodies in serum) Review of other AEs and concomitant medications

[0163] Follow-up (Day 57 [± 3 days]) Participants will return to the CRU for follow-up on Day 57 (28 days ± 3 days after Visit 6).

[0164] At the follow-up visit, the following steps are performed: · Urine pregnancy test (WOCBP only) Urine testing Vital signs (systolic and diastolic blood pressure, respiratory rate, pulse rate, oral temperature) (after participants had rested in a supine position for at least 5 minutes) ·Intensive physical examination ·NP swabs for evaluation of vaccine shedding and humoral immunogenicity (IgA antibodies) Blood sampling for clinical tests (hematology, biochemistry) Blood sampling to assess immunogenicity (antibodies in serum) Review of other AEs and concomitant medications 12-lead ECG (after participants rested in supine position for at least 5 minutes)

[0165] Follow-up (Day 113 [± 10 days]) Participants will return to the CRU for follow-up on Day 113 (Day 84 ± 10 days after Visit 6).

[0166] At the follow-up visit, the following steps are performed: · MAE, NCI, and SAE only reviews -Review of concomitant medications of particular interest ·NP swabs for vaccine shedding assessment Blood sampling to assess immunogenicity (antibodies in serum)

[0167] End of clinical trial (209th day [± 10 days]) Participants will return to the CRU for EOS follow-up on day 209 (6 months ± 10 days after visit 6).

[0168] At the follow-up visit, the following steps are performed: · Urine pregnancy test (WOCBP only) Blood sampling to assess immunogenicity (antibodies in serum) · MAE, NCI, and SAE only reviews -Review of concomitant medications of particular interest. This visit will mark the final participation in this study.

[0169] Immunogenicity assessment Collection and analysis of serum samples Blood samples for serum will be collected, processed, and shipped for analysis according to the institution's standard operating procedures and clinical laboratory manuals prior to dosing and at the time points specified in the evaluation schedule. Serum will be separated within 2 hours and frozen at -80°C (± 10°C).

[0170] Serum is shipped frozen to Codagenix laboratories (Farmingdale NY USA) and tested as follows:

[0171] RSV immunoglobulin G RSV-specific serum IgG antibodies are analyzed by ELISA using RSV-A2 whole virus lysate as the target antigen. Briefly, heat-inactivated serum samples are serially diluted and reacted with assay wells coated with RSV antigen. Bound RSV-specific serum antibodies are detected by colorimetric detection using horseradish peroxidase (HRP) and a secondary anti-human IgG antibody conjugated to soluble o-phenylenediamine dihydrochloride (OPD) as the colorimetric substrate. The amount of bound RSV-specific serum antibodies is calculated based on a standard curve of purified human IgG standards run in parallel with each assay. RSV-specific IgG antibody titers are reported as concentrations in ng / mL serum. Further details are outlined in the Clinical Laboratory Manual.

[0172] RSV neutralizing antibody RSV-specific serum neutralizing antibodies are analyzed by a microneutralization assay using RSV-A2 virus as target. Briefly, heat-inactivated serum samples are serially diluted and reacted with 100×TCID50 of RSV virus in tissue culture 96-well plates for 1 h at 37°C. 20,000 Vero cells are added to each well as a virus substrate. 72 h after infection, assay wells are fixed and stained by immunohistochemistry using RSV mouse monoclonal antibody 2F7 (Novus Biological) and secondary goat anti-mouse IgG conjugated to HRP and soluble OPD as colorimetric substrates. RSV neutralization titers are reported as the reciprocal of the dilution of serum that results in a 50% reduction in color development [i.e., (uninhibited virus control-uninfected cell control) / 2 signal]. Further details are outlined in the Clinical Laboratory Manual.

[0173] Isolation and analysis of peripheral blood mononuclear cells PBMC blood samples will be collected at the time points specified in the evaluation schedule and sent for processing and analysis as outlined in the clinical laboratory manual for the ELISpot assay.

[0174] Upper respiratory tract specimen collection and analysis Nasopharyngeal swabs from both nostrils will be collected at the time points indicated in the assessment schedule and processed and sent for analysis as outlined in the clinical laboratory manual. These samples will be analyzed for vaccine and RSV-specific IgA.

[0175] Immunoglobulin A RSV-specific IgA antibodies are analyzed by ELISA using RSV-A2 whole virus lysate as the target antigen. Briefly, NP swabs in Universal Viral Transport Medium (UVTM) are serially diluted and reacted with assay wells coated with RSV antigens. Bound RSV-specific serum antibodies are detected by colorimetric detection using a secondary anti-human IgA antibody conjugated to HRP and soluble OPD as the colorimetric substrate.

[0176] The amount of bound RSV-specific serum antibody is calculated based on a standard curve of purified human IgA standards run in parallel with each assay. RSV-specific IgG antibody titers are reported as concentrations in ng / mL serum. Further details are outlined in the Clinical Laboratory Manual.

[0177] Symptomatic Respiratory Viral Infections Study Participants with moderate to severe illness consistent with a viral respiratory infection, in the opinion of the investigator, will have an NP swab or other appropriate specimen collected for a multiplex PCR panel of respiratory viruses. This can be obtained by NP swab specimen or (especially if the subject is at an outside facility) other standard specimens such as sputum or nasal wash. All RSV-positive samples will be frozen and sent to Codagenix for sequence analysis. Swabs will be collected, processed, and shipped for analysis as outlined in the clinical laboratory manual.

[0178] This testing will be performed at the investigator's discretion and may be in addition to routine monitoring of vaccine virus shedding.

[0179] Vaccine shedding by PCR Nasopharyngeal swabs are collected. The presence and magnitude of RSV shedding in NP secretions is assessed at the sponsor's laboratory using the ARIES Flu A / B & RSV Assay (Luminex Corporation, Ref#50-10020) performed on the Luminex ARIES diagnostic platform. For the purposes of RSV shedding quantification, the ARIES Flu A / B & RSV Assay is modified as a "trial use only" test by calculating the amount of virus present in the sample based on a standard curve of known amounts of RSV reference virus. Shedding titers are reported as FFU equivalents / mL (FFUeq / mL). After each vaccination, if two consecutive samples are below detection, subsequent samples are not tested. Further details are outlined in the Clinical Laboratory Manual.

[0180] Sequence analysis of the shedding virus Isolates obtained during episodes of symptomatic RSV infection and the last positive shedding samples after each vaccination will be sequenced by next generation sequencing after conversion of viral RNA to cDNA using commercially available reverse transcriptase PCR kits. Details are outlined in the Clinical Laboratory Manual.

[0181] RSV viremia by PCR The presence of CodaVax-RSV in serum (viremia) will be assessed at the sponsor's laboratory using the Luminex assay described herein. Serum samples that are positive by PCR will be confirmed by viral culture. Further details are outlined in the Clinical Laboratory Manual.

[0182] Reactogenicity After vaccination, participants will be provided with a diary card and a thermometer to record oral temperature and involuntary reactogenic events and severity for 7 days after each vaccination.

[0183] Events recorded on the diary cards will be considered as non-spontaneous local or non-spontaneous systemic events and will be graded in accordance with the FDA Guidance for Industry: Toxicity Grading Scale for Healthy Adult and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical Trials.

[0184] Site staff will review the diary with the subject to ensure it is clear and that ratings are consistent with the toxicity grading scale provided. Site staff may ask the subject to adjust ratings if indicated.

[0185] Diary cards will be issued to participants at Visit 2 (Day 1) and collected at Visit 4 (Day 8). Diary cards will be issued again at Visit 6 (Day 29) and collected at Visit 8 (Day 36). If the event continues beyond Day 8 or Day 36, site staff will record the study date the symptom ended on the source document and enter this information into the Reactogenic Event / Symptom CRF.

[0186] NOTE: Involuntary reactogenic events are not recorded as AEs unless they are severe (i.e., they meet the criteria for an SAE).

[0187] NOTE: If two or more participants experience similar grade 3 events, the MM will be notified.

[0188] Involuntary localized events Local reactogenic symptoms collected are: runny nose, stuffy nose, nasal irritation / pain, sneezing, coughing, sore throat, sore eyes, sore ears, changes in vision, changes in smell, changes in taste.

[0189] Involuntary systemic events Systemic reactogenic symptoms collected are: headache, fatigue, myalgia, arthralgia, chills, sweating, vomiting, diarrhea, chest pain / pleurisy, shortness of breath.

[0190] heat Body temperature (oral) should be measured daily by self-assessment and recorded on a diary card from each vaccination up to 7 days after vaccination. Prophylactic administration of antipyretics is not permitted for 4 hours before vaccination and for 72 hours after vaccination, however subjects may use over-the-counter antipyretics and / or analgesics to treat symptoms. Symptoms and temperature should be recorded at least 6 hours after the last administration of such medication.

[0191] immunogenicity Immunogenicity parameter values ​​are listed by participant and summarized by dose level (CodaVax-RSV low dose, high dose, and placebo) using the PP population. If more than 10% of participants at any dose level were excluded from the PP population, the ITT population is also presented. For RSV-specific IgG (in serum) and IgA (NP swab specimen) antibodies measured by ELISA, GMT and GMR by time point are modeled using a linear mixed-effects model with participant as a random effect, time point (random) and treatment group as fixed effects, and baseline titer as a covariate. The proportion of participants who seroconvert is expressed as N (%), and 95% Wilson confidence intervals are reported for each group. For neutralizing antibodies measured in serum, GMT and GMR by time point are modeled using the same linear mixed-effects model as for RSV-specific IgG above. Seroconversion rates at each time point and for each treatment group are displayed with 95% CI. Scatter plots of baseline RSV neutralizing antibody titers and post-vaccination immunogenicity measures are presented. T cell responses to RSV / A will be listed by participant and summarized by treatment and protocol specified time points. Subgroup analyses of immunogenicity measures with low and high baseline RSV neutralizing antibody titers will be completed. No adjustment for multiple comparisons will be applied.

[0192] Example 2 The following example is based on data collected in the clinical trial described in Example 1 above.

[0193] Data from the ELISpot performed by 360 Bio were reanalyzed. Detailed conditions and planned analyses are described in the 360 ​​Bio laboratory assay report and statistical analysis plan. In this post-hoc analysis, only values ​​from the 1:10 stimulation of the low-dose cohort (13 vaccinees and 7 placebo recipients) are included.

[0194] In a post-hoc analysis, the change from baseline (day 1) to the highest value (peak) at any time point after vaccination was evaluated, and subjects with a ≥1.3-fold increase from baseline were defined as responders.

[0195] The baseline and peak values of vaccinated and placebo - vaccinated individuals were compared using a paired t - test. The fold change from baseline to peak after vaccination was calculated by substituting a value of <LLOQ(50 SFU) as 49 SFU. The Mann - Whitney test was used to compare the fold changes between the vaccine group and the placebo group.

[0196] Results: Total number of detectable RSV - specific responses: 10 / 13 of the vaccinated volunteers showed a detectable increase in the number of spot - forming units compared to day 1, indicating the occurrence of a T - cell response (76.9%). In the placebo group, an increase was seen in only 1 / 7 (14.2%) of the volunteers. The baseline and peak values of the vaccine group were statistically different in the paired t - test (p < 0.01), while there was no difference in the placebo group (p = 0.39). The SFU values at baseline and peak for both placebo - and CodaVax - RSV - vaccinated volunteers are shown in Figure 1A, and the individual changes are shown in Figure 1B.

[0197] Furthermore, in 6 / 13 (46%) of the CodaVax - vaccinated volunteers, an increase more than 1.3 - fold in the number of SFU was seen compared to 1 / 7 (14.3%) of the placebo - vaccinated individuals. The fold change in the number of SFU between the peak day and day 1 is shown in Figure 2.

[0198] Volunteers vaccinated with CodaVax - RSV showed an average 1.5 - fold increase in the number of SFU, while the group treated with placebo had a - 0.98 - fold change. As evaluated by the Mann - Whitney test, the difference in the increase in SFU between the placebo group and the CodaVax - RSV - vaccinated group was statistically significant (p < 0.03).

[0199] A post-hoc examination of this ELISpot data identified a statistically significant increase in cellular immune responses after vaccination, which was not immediately apparent in the planned analyses, which were limited to evaluation from baseline to each post-vaccination time point, rather than from baseline to the peak post-vaccination time point. Furthermore, the planned analyses did not include a definition of responders as was done here.

[0200] Future trials may use a 1.3-fold increase to define responders and also plan to process PBMCs from whole blood within a shorter time frame from collection, which may allow for better detection of activated lymphocytes, which are fragile and can be lost during transport.

[0201] Example 3 The following example is also based on data collected in the clinical trial described in Example 1 above.

[0202] RSV-specific IgA versus total IgA antibodies measured by ELISA in nasopharyngeal swabs at presentation. Results are presented below. TIFF2024513917000006.tif163165

[0203] Example 4 MinL-4.0 (RSV-MinL NPM2-1(N88K)L, Le Nouen et al, 2017) (SEQ ID NO: 1) TIFF2024513917000007.tif76157TIFF2024513917000008.tif241158TIFF20245139170 00009.tif241157TIFF2024513917000010.tif241156TIFF2024513917000011.tif110154

[0204] The four nucleotide differences in RSV-MinL NPM2-1(N88K)L (MinL4.0) compared to the MinL parent (introduced by reverse genetics, see Le Nouen 2017) are A1547G, A2687T, C7758A, and C11883T. TIFF2024513917000012.tif101128TIFF2024513917000013.tif24255TIFF2024513917000014.tif115128

[0205] Example 5 T cell response The following example is also based on data collected in the clinical trial described in Example 1 above.

[0206] No differences were found between CodaVax-RSV dose groups or the overall CodaVax-RSV group and placebo in planned analyses of post-dose T cell responses (geometric mean SFU) measured by RSV-stimulated PBMCs in either analysis population. GMFRs ranged from 0.9 to 1.3.

[0207] In a post-hoc analysis defining responders as participants with an increase from baseline to peak values, the overall responder rate in the CodaVax-RSV group was 66.7% (95% CI: 46.7, 82.0%) compared with 16.7% (95% CI: 4.7, 44.8%) for placebo.

[0208] The analysis was repeated using the raw data (rather than substituting a value below the LLOQ as the LLOQ, which would mask changes in SFU<50). In this analysis, participants who received Coda Vax-RSV had increased SFUs, but no change in placebo recipients. As shown in Figure 3, there was a potential dose effect in this response. The GMFR on day 8 using 1 / 10 viral lysate stimulation was 2 × 10 6 In the PFU group, it was 3.6 (95% CI: 1.5, 8.7), and 2 × 10 5The GMFR was 1.4 (95% CI: 0.9, 2.2) in the PFU group and 2.1 (95% CI: 1.3, 3.4) in the overall CodaVax-RSV group, but was unchanged in the placebo group (GMFR: 0.9 [95% CI: 0.4, 2.1]). Differences were similar at other time points and for 1 / 80 viral lysate (Figure 3).

[0209] A cellular immune response was observed after the first dose.

[0210] In Figure 3, the vertical dotted line (1-fold change) means no change. The lines around the point estimates indicate the 95% confidence interval. Thus, the fact that the confidence intervals across the high dose and active groups do not exceed 1 indicates that there is a significant increase in T cell responses. The placebo group was unchanged, with confidence intervals on either side of the dotted line. There was a trend for an increase in the low dose group, but the fact that the confidence interval fell below 1 indicates that it was not statistically significant.

[0211] Various embodiments of the present invention are described in the above detailed description. Although these descriptions directly describe the above embodiments, it should be understood that those skilled in the art may envision modifications and / or variations to the specific embodiments shown and described herein. Any such modifications or variations that fall within the scope of this description are also intended to be included within the scope. Unless otherwise noted, it is the intention of the inventors that the words and phrases in this specification and claims be given their ordinary and customary meaning to those skilled in the applicable technical field(s).

[0212] The foregoing description of various embodiments of the invention known to the applicant at the time of filing this application has been presented and is intended for purposes of illustration and description. This description is not intended to be exhaustive or to limit the invention to the precise form disclosed, as many modifications and variations are possible in light of the above teachings. The described embodiments illustrate the principles of the invention and its practical application, enabling one skilled in the art to utilize the invention in various embodiments, with various modifications suited to the particular applications intended. Therefore, it is not intended that the invention be limited to the specific embodiments disclosed for carrying out the invention.

[0213] While specific embodiments of the present invention have been shown and described, it will be apparent to those skilled in the art that, based on the teachings herein, changes and modifications may be made without departing from the present invention and its broader aspects, and therefore, the appended claims are intended to encompass within their scope all changes and modifications as are within the true spirit and scope of the present invention. In general, it will be understood by those skilled in the art that the terms used herein are generally intended as "non-limiting" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," and the term "including" should be interpreted as "including but not limited to").

[0214] Unless otherwise stated, the terms "a" and "an" and "the" and similar references used in the context of describing specific embodiments of the present application (particularly in the context of the claims) can be construed to cover both the singular and the plural. Recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value falling within the range. Unless otherwise indicated herein, each separate value is incorporated herein as if it were individually recited herein. All methods described herein may be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "such as") provided with respect to specific embodiments herein is intended only to better illustrate the application and does not pose a limitation on the scope of the application as otherwise claimed. The abbreviation "eg" is derived from the Latin exempli gratia and is used herein to indicate a non-limiting example. Thus, the abbreviation "eg" is synonymous with the term "for example." No language in the specification should be construed as indicating any non-claimed element essential to the practice of the application.

[0215] "Optional" or "optionally" means that the subsequently described circumstance may or may not occur, and that the description includes instances where the circumstance occurs and instances where it does not occur.

[0216] Groupings of alternative elements or embodiments of the disclosure disclosed herein are not to be construed as limiting. Each group member may be referenced and claimed individually or in any combination with other members of the group or other elements described herein. One or more members of a group may be included in or deleted from a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the modified group and thus fulfills the written description of all Markush groups used in the appended claims.

Claims

1. A pharmaceutical composition for inducing an immune response in a subject, comprising a deoptimized respiratory syncytial virus (RSV), The pharmaceutical composition is used such that one or more doses of an effective amount of the pharmaceutical composition are administered intranasally to the subject; The effective amount is about 10 ３ ~10 ９ the deoptimized RSV in focus forming units (FFUs); The immune response comprises an IgA immune response, a cellular immune response, or both. The pharmaceutical composition.

2. 10. The pharmaceutical composition of claim 1, wherein intranasally administering one or more doses of the effective amount of the pharmaceutical composition comprises administering two doses, the second dose being administered about 28 days after the first dose, at least 28 days after the first dose, or about 28-35 days after the first dose.

3. 3. The pharmaceutical composition of claim 1 or 2, wherein administering intranasally comprises administering via nasal drops or nasal spray.

4. The effective amount is about 10 ５ 2. The pharmaceutical composition of claim 1, wherein the concentration of the recombinant human leukocyte antigen (LEA) is about 10 FFU, about 2x10 5 FFU, about 10 6 FFU, or about 2x10 6 FFU.

5. and / or wherein said effective amount is provided in a volume of about 1 mL; and / or said effective dose being administered in about four aliquots; The pharmaceutical composition of claim 1.

6. The effective amount of a single dose is administered in about four aliquots; and (a) a first aliquot is administered to a first nostril and a second aliquot is administered to a second nostril within 10 minutes of said first aliquot being administered to said first nostril; and about 5-20 minutes after the second aliquot is administered to the second nostril, a third aliquot and a fourth aliquot are administered in any order to the first nostril and the second nostril; or (b) a first aliquot is administered to a first nostril and a second aliquot is administered to a second nostril within 5 minutes of the first aliquot being administered to the first nostril; and about 10-15 minutes after the second aliquot is administered to the second nostril, a third aliquot and a fourth aliquot are administered in any order to the first nostril and the second nostril; The pharmaceutical composition according to claim 5. (i) the cellular immune response comprises the generation of a T cell response; or (ii) the cellular immune response comprises the development of a T cell response, and the development of the T cell response is compared to a baseline level of response based on a placebo group or compared to a time point prior to vaccination of the subject; The pharmaceutical composition of claim 1.

8. 2. The pharmaceutical composition of claim 1, wherein enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.3-fold, at least a 1.4-fold, at least a 1.5-fold, at least a 1.6-fold, or at least a 1.7-fold increase in spot forming units (SFU) between baseline and highest value at the post-vaccination time point.

9. The effective amount is about 10 ６ 2. The pharmaceutical composition of claim 1, wherein the vaccine comprises at least about 10 FFU, about 2x10 FFU, about 10 FFU, or about 2x10 FFU, and wherein eliciting an immune response comprises a statistically significant increase in T cell responses compared to a pre-vaccination time point. (i) the effective amount is about 10 ６ FFU, and eliciting the immune response comprises at least a 2.4-fold increase in IgA levels in the subject. (ii) the effective amount is about 2×10 6 FFU and eliciting an immune response comprises at least a 2.4-fold increase in IgA levels in the subject; (iii) the effective amount is about 10 5 FFU and eliciting the immune response comprises at least a 1.1-fold increase in IgA levels in the subject; (iv) the effective amount is about 2×10 5 FFU and eliciting an immune response comprises at least a 1.1-fold increase in IgA levels in the subject; (v) the effective amount is about 10 5 to 10 6 FFU and eliciting the immune response comprises at least a 1.3-fold increase in IgA levels in the subject; or (vi) the effective amount is about 2×10 5 to 2×10 6 FFU and eliciting an immune response comprises at least a 1.3-fold increase in IgA levels in the subject; The pharmaceutical composition of claim 1.

11. The deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:

1. The pharmaceutical composition of claim 1.

12. The object is (i) You are under 18 years of age; (ii) is under 13 years of age; (iii) is under 7 years of age; (iv) Under 2 years of age; (v) Are between 18 and 49 years of age; or (vi) is at least 50 years of age; The pharmaceutical composition of claim 1.

13. A pharmaceutical composition for inducing a T cell immune response in a subject, comprising a deoptimized respiratory syncytial virus (RSV), said pharmaceutical composition is used such that an effective amount of a first dose of said pharmaceutical composition is administered intranasally to said subject, and about 28-35 days after said first dose, said effective amount of a second dose of said pharmaceutical composition is administered intranasally; The effective amount is about 10 ６ Or about 2 x 10 ６ the deoptimized RSV of focus forming units (FFUs), wherein the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting the T cell immune response comprises an increase in the T cell response compared to a pre-vaccination time point. The pharmaceutical composition.

14. A pharmaceutical composition for inducing a T cell immune response in a subject, comprising a deoptimized respiratory syncytial virus (RSV), said pharmaceutical composition is used such that an effective amount of a first dose of said pharmaceutical composition is administered intranasally to said subject, and about 28-35 days after said first dose, said effective amount of a second dose of said pharmaceutical composition is administered intranasally; The effective amount is about 10 ５ Or about 2 x 10 ５ the deoptimized RSV of focus forming units (FFUs), wherein the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting the T cell immune response comprises an increase in the T cell response compared to a pre-vaccination time point. The pharmaceutical composition.

15. A pharmaceutical composition for inducing an IgA immune response, a cellular immune response, or both in a subject, comprising a deoptimized respiratory syncytial virus (RSV), said pharmaceutical composition is used such that an effective amount of a first dose of said pharmaceutical composition is administered intranasally to said subject, and about 28-35 days after said first dose, said effective amount of a second dose of said pharmaceutical composition is administered intranasally; The effective amount is about 2×10 ５ the deoptimized RSV for focus forming units (FFU), wherein the deoptimized RSV comprises a genome or antigenome having the sequence of SEQ ID NO:1, and wherein enzyme-linked immunosorbent spot (ELISpot) analysis of peripheral blood mononuclear cells (PBMCs) from the subject shows at least a 1.3-fold increase in spot forming units (SFU) between baseline and highest value at a post-vaccination time point; or The effective amount is about 2×10 ６ the deoptimized RSV in focus forming units (FFUs), the deoptimized RSV comprising a genome or antigenome having the sequence of SEQ ID NO:1, and eliciting an immune response comprises at least a 2.4-fold increase in IgA levels in the subject. The pharmaceutical composition.

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